CN110305879A - A kind of southern corn leaf blight ChCDC3 gene and its application - Google Patents
A kind of southern corn leaf blight ChCDC3 gene and its application Download PDFInfo
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- CN110305879A CN110305879A CN201910618994.6A CN201910618994A CN110305879A CN 110305879 A CN110305879 A CN 110305879A CN 201910618994 A CN201910618994 A CN 201910618994A CN 110305879 A CN110305879 A CN 110305879A
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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
Abstract
A kind of southern corn leaf blight ChCDC3 gene and its apply microorganism belonging to genus gene engineering technology field, the ChCDC3 gene that control bacterium colony growth rate, illumination and ascospore provided by the invention from southern corn leaf blight is formed, DNA sequence dna is as shown in SEQ ID No:1；The protein of the ChCDC3 coded by said gene of offer, amino acid sequence is as shown in SEQ ID No:2；ChCDC3 gene can be applied in Genes For Plant Tolerance corn southern leaf blight genetic engineering field；It lacked, be mutated or modified by controlling the protein C hCDC3 that bacterium colony growth rate, conidium and ascospore are formed to southern corn leaf blight, and it is limited to form its slow growth, conidium and ascospore, southern corn leaf blight Life Cycles are caused to be obstructed, it can be used as target to apply in designing and screening anti-corn southern leaf blight medicament, the proteinoid is not present especially in plant, to plant safety.
Description
Technical field
The invention belongs to technical field of microbial genetic engineering, and in particular to epiphyte pathogenic is controlled in plant protection art
The discovery of new gene and its application of coding protein.
Background technique
The Invisible element scientific name of southern corn leaf blight are as follows: Bipolaris maydis belongs to the concordant Helminthosporium of Deuteromycotina,
Its Perfect stage scientific name is Cochliobolus heterostrophus, belongs to the different cochliobolus of Ascomycotina.Nineteen twenty-five is made
It is found for the first time for the pathogenic bacteria of maize diseases.Southern corn leaf blight belongs to filamentous fungi, when mycelia grows into certain phase
Afterwards, under the influence of external environmental condition and oneself factor, conidium is grown on conidiophore top or side.Point
Raw spore has diaphragm 3-13, wherein most of a with diaphragm 7-9, dark brown, oblong, both ends blunt circle, multidirectional one
Lateral bend, intermediate thick, both sides are thin, and size is 10-17 μm of 30-115 μ m, and omphalion is depressed within basal cell.Conidium is more
It is sprouted from two teloblasts and grows germ tube, intermediate cell can also sprout germ tube out sometimes.The temperature range of illumination is 15
~33 DEG C, optimum temperature range is 23~25 DEG C；The temperature range of conidia germination is 5~42 DEG C, and optimum temperature range is
26~32 DEG C.The ability that conidium resists drying is stronger, can survive at least a year on corn seed.Conidium is wet
Under conditions of profit, it can sprout and grow germ tube, mycelia is formed by apical growth.
Under natural conditions, mycelia grows into certain phase, under external environmental condition, it is also possible to enter zoogamy rank
Section, the zoogamy stage of southern corn leaf blight be it is heterothallic, have on its genome a single site with it is sexual
Reproduction is related, is named as MAT-1, and there are two types of different forms, i.e. MAT-1-1 and MAT-1-2 in the site.Southern corn leaf blight
The zoogamy stage it is uncommon, can find the shell of ascus in withered diseased tissues once in a while.It can pass through in laboratory conditions
Artificial induction generates the shell of ascus, and the shell of ascus of southern corn leaf blight is referred to as " pseudothecium ".The shell of ascus is from forming maturation
One month time is taken around, after the mature shell of ascus meets water, top rupture releases ascus and ascospore.The shell of ascus
Black, spherical, rostral is obvious, and length is embedded in host's diseased tissues, and surface can grow mycelium and conidiophore；It is internal raw close
Barrel-shaped ascus.Ascus top blunt circle, base portion have handle.8 are formed by meiosis and mitosis in mature ascus
Linear monoploid ascospore, ascospore mutually wind spiral in ascus, and when sprouting, each cell can be grown
Germ tube, and then grow up to mycelia.Studies have found that in mature ascus 80% have complete 8 ascospores, 20% have 4-
7 ascospores.Southern corn leaf blight relies primarily on conidium with the biography of air-flow and rainwater with popular in infecting for field
It broadcasts, if it is possible to control conidial formation, that will substantially reduce source of infection, reduce the generation of corn southern leaf blight, improve beautiful
Rice yield.
Corn southern leaf blight is a kind of main Maize Leaf fungal disease, occurs mainly in the corn producing region of warm moist.
In the 1970s, due to a large amount of plantations of the corn containing T-type male sterile cytoplasm (T-cms), so that corn southern leaf blight
It is very popular in the U.S., causes 16,500,000,000 kilograms of the corn underproduction, account for the 15% of american corn total output, about 1,000,000,000 beauty of the loss output value
Member, because caused loss has been more than to occur to be very popular in the European late blight of potato in 1840 and shock the whole world.Early in
The 1920s, just there was the generation of corn southern leaf blight in the Jiangsu Province of China, but only occurred in the rainy time, and mostly
It is popular in the later period of corn growth, seldom cause serious economic loss.But due in the 1960s, corn is susceptible miscellaneous
The large area plantation for handing over kind, so that the harm of helminthosporium maydis is on the rise, becomes main maize leaf disease.To 20th century 60
Mid-nineties 90 causes Hebei and Hubei some areas Severe Reduction due to the serious generation of corn southern leaf blight, and general plot subtracts
Production has reached 20% or more, and the serious plot underproduction is up to 80%, or even total crop failure.After the 1970s, corn is disease-resistant
The popularization of kind, helminthosporium maydis have obtained basic control, but due to the plantation large area unification of disease-resistant variety with
And global air temperature warming, in certain corn producing regions in China, there is serious generation in when generation of helminthosporium maydis, causes heavy loss.
Southern corn leaf blight conidium and ascospore formation are furtherd investigate, identify the pass of southern corn leaf blight sporogenesis
The key factor not only facilitates and discloses the pathogenic molecular mechanism of this necrotrophic disease fungus of southern corn leaf blight, also may be used
The protein of fungicide action target can be found to can be used as, therefrom for exploitation control of leaf spot of corn and other similar diseases
Efficient medicament establish theory and technology basis.
CDC3 is the gene of a unknown function in southern corn leaf blight.By analyzing southern corn leaf blight ChCDC3 base
The function of cause evaluates effect of the gene in southern corn leaf blight growth and development process, is conducive to identify potential prevention and treatment target
Mark, for screening the medicament of novel prevention and control southern corn leaf blight.
Summary of the invention
The purpose of the present invention is intended to provide the gene and its coding of a kind of control fungus conidium and ascospore formation
Protein.
The gene source that control conidium provided by the present invention and ascospore are formed is in southern corn leaf blight, title
For ChCDC3, DNA sequence dna is as shown in SEQ ID No:1.The DNA sequence dna is the open reading frame of ChCDC3 gene, by 1403
A nucleotide composition, wherein including 2 intron sequences.
The present invention provides southern corn leaf blight (Cochliobolus heterostrophus) ChCDC3 coded by said gene
Protein, as shown in SEQ ID No:2, which is made of amino acid sequence 432 amino acid.
The ChCDC3 gene that control conidium and ascospore from southern corn leaf blight are formed can be applied to plant
Anti- corn southern leaf blight genetic engineering field.
To the egg of the ChCDC3 coded by said gene of control conidium and ascospore formation from southern corn leaf blight
White matter is lacked, is mutated or modified, and forms its conidium and ascospore and defect occurs, can be used as target and designing
It is applied with screening in anti-corn southern leaf blight medicament, the protein is not contained especially in plant, so safer to plant.
Present invention demonstrates that the missing of southern corn leaf blight (Cochliobolus heterostrophus) ChCDC3 gene
Or mutation, cause southern corn leaf blight illumination to be remarkably decreased, and normal ascospore cannot be formed, illustrates jade
Rice stigma germ (Cochliobolus heterostrophus) ChCDC3 gene is must in southern corn leaf blight life cycle
The gene needed.Currently, influencing to be not yet reported that in terms of ascospore is formed about CDC3 gene, this research is to report for the first time.
Therefore, screening can prevent the compound of the expression, modification and positioning of the gene expression and its protein, can effectively control jade
The generation of rice helminthosporium maydis, to help to develop new type bactericide, i.e., southern corn leaf blight provided by the present invention
One important use of (Cochliobolus heterostrophus) ChCDC3 gene is: the expression of the gene is encoded with it
Protein product expression, modification and positioning, can be used as important candidate targets site, be used for anti-southern corn leaf blight medicament
Design and screening.
Detailed description of the invention
Fig. 1 is that the structural domain of ChCDC3 protein predicts schematic diagram
Wherein: one conservative CDC-Septin functional domain of discovery；
Fig. 2 is the schematic diagram of southern corn leaf blight ChCDC3 gene knockout strategy (carrying out gene replacement by homologous recombination)
Wherein: ChC4 is southern corn leaf blight wild-type strain；Δ Chcdc3 is the deletion mutant of ChCDC3 gene；Draw
Object F1/R1 and F2/R2 is respectively used for amplifying the upstream and downstream sequence of ChCDC3 gene, the homology arm as knockout；Primers F/R, U/
NLC37, NLC38/D are for verifying mutant；
Fig. 3 is that the PCR of southern corn leaf blight ChCDC3 deletion mutant body verifies electrophoretogram
Wherein: F/R, U/NLC37, D/NLC38 are the primer；1 is southern corn leaf blight wild-type strain, and 2 and 3 are
ChCDC3 deletion mutant body；It (1) is part ChCDC3 gene magnification result；(2) add portion for ChCDC3 gene upstream sequence
Partial tide mycin sequence amplification result；(3) add part hygromycin sequence amplification result for ChCDC3 downstream of gene sequence；
Fig. 4 is the deletion mutant and wild-type strain and complementing strain Δ of southern corn leaf blight ChCDC3 gene
The cultural characteristic of ChCDC3-C compares photo；
Wherein: used medium CMX, 24 DEG C of cultures, observation in 7 days is taken pictures after inoculation；WT is that southern corn leaf blight is wild
Type bacterial strain, other strain number meanings are as described above.
Fig. 5 is the deletion mutant and wild-type strain and complementing strain Δ of southern corn leaf blight ChCDC3 gene
Colony diameter after ChCDC3-C culture 7 days；
Wherein: used medium CMX, 24 DEG C of cultures measure for 7 days after inoculation；WT is southern corn leaf blight wild-type bacteria
Strain, other strain number meanings are as described above.
Fig. 6 is the deletion mutant and wild-type strain and complementing strain Δ of southern corn leaf blight ChCDC3 gene
The microcosmic comparison picture of ChCDC3-C conidium growth
Wherein: spore used produces spore in culture 7-9d on CMX culture medium by corresponding strain inoculated, prepares spore suspension
Liquid is added dropwise on glass slide, and observation is taken pictures under the microscope.
Fig. 7 is the deletion mutant and wild-type strain and complementing strain Δ ChCDC3-C conidium phase of ChCDC3 gene
To yield；
Wherein: spore used produces spore in culture 9d on CMX culture medium by corresponding strain inoculated, prepares spore suspension
Liquid calculates spore concentration using blood cell plate counter.
Fig. 8 is that the deletion mutant of ChCDC3 gene is sprouted with wild-type strain and complementing strain Δ ChCDC3-C conidium
Hair rate.
Wherein: spore used produces spore in culture 9d on CMX culture medium by corresponding strain inoculated, prepares spore suspension
It is added dropwise on glass slide, 24 DEG C of moisturizing culture 6h, measures spore germination rate.
Fig. 9 is that the deletion mutant of ChCDC3 gene and wild-type strain and the complementing strain Δ ChCDC3-C shell of ascus generate
Quantity.
Wherein: the deletion mutant of ChCDC3 gene and wild-type strain and complementing strain Δ ChCDC3-C and corn is small
Pinta bacteria strain CB7 is hybridized, and the quantity of the shell of ascus is measured after downloading in 25 DEG C and cultivate 21d on Sach culture medium.
Figure 10 is the deletion mutant and wild-type strain and complementing strain Δ ChCDC3-C ascospore of ChCDC3 gene
Microexamination picture.
Wherein: the deletion mutant of ChCDC3 gene and wild-type strain and complementing strain Δ ChCDC3-C and corn is small
Pinta bacteria strain CB7 is hybridized, and obtains ascospore and under the microscope after cultivating 21d on 25 DEG C of downloading Sach culture mediums
Observation is taken pictures.
Specific embodiment
It in order to better describe the present invention, is further described below by embodiment, the method in following embodiments,
It unless otherwise instructed, is conventional method.
Corn southern leaf blight bacterial strain used in the present invention (Cochliobolus heterostrophus) is jade of falling ill from field
It is collected on rice.
The correlation analysis of 1 ChCDC3 gene of embodiment
Southern corn leaf blight ChCDC3 gene is that this team is compared in southern corn leaf blight using the CDC gene in saccharomycete
To acquisition.The open reading frame of southern corn leaf blight ChCDC3 gene is made of 1403 nucleotide, includes 2 intrones.It compiles
The protein product of code is made of 432 amino acid, structure domain analysis discovery, the protein of southern corn leaf blight ChCDC3 coding
Include a conservative CDC-Septin functional domain (see Fig. 1).
The knockout of 2 southern corn leaf blight ChCDC3 gene of embodiment
1) amplification of ChCDC3 gene upstream and downstream and hygromycin gene
Using primers F 1 (5'-T C G T G C C T C C T C A T T T A A C C-3') and R1 (5'-TC
CTGTGTGAAATTGTTATCCGCTGGAGAAGTAGCACCGTTGGA-3'), with the base of southern corn leaf blight wild-type strain C4
Because group DNA is template amplification ChCDC3 upstream region of gene 942p segment, using F2 (5'-GTCGTGACTGGGAAAACCCTGGCGGC
TATCGGTAAACGGGACAA-3') southern corn leaf blight ChCDC3 is expanded with R2 (5'-CCCGCTCATGATACTCTTCG-3')
Downstream of gene 805bp segment, using primer M13F (5'-CGCCAGGGTTTTCCCAGTCACGAC-3') and M13R (5'-
AGCGGATAACAATTTCACACAGGA-3'), using carrier pUCATPH as template amplification 2549bp hygromycin gene.Reaction system
Are as follows: 10mmol/L dNTP Mixture, 1 μ L；5 × PCRbuffer, 10 μ L；Each 2.5 μ L of upstream and downstream primer (10 μm of ol/mL)；
Template DNA, 2 μ L；Phusion polymerase, 0.5 μ L (5U)；ddH2O, 31.5 μ L；Amplification program are as follows: 98 DEG C of initial denaturations 2
Minute, it then (1) 98 DEG C, is denaturalized 20 seconds；It (2) 65 DEG C, anneals 30 seconds；(3) 72 DEG C, extend 30 seconds；(4) it recycles 30 times；(5)72
DEG C extend 10 minutes.Above-mentioned 3 segments are transferred to jointly in southern corn leaf blight wild-type strain C4.
2) southern corn leaf blight converts
A. the production spore culture of southern corn leaf blight
A small amount of southern corn leaf blight C4 bacterial strain conidium is taken from -80 DEG C of refrigerators, is added dropwise in [the every liter of CMX training of CMX culture medium
Feeding base includes: 0.1g/mL calcium nitrate tetrahydrate solution 10mL, 10mL solution B, 0.5mL trace element solution, and 1g yeast extracts
Object, 0.5g digest casein, 0.5g acidolysis casein, 10g xylose, 20g agar powder.(every liter of solution B includes: 20g biphosphate
Potassium, 25g bitter salt, 15g sodium chloride) (every liter of trace element solution includes: 57.2mg boric acid, five hydrated sulfuric acid of 393mg
Copper, 13.1mg potassium iodide, 60.4mg Manganous sulfate monohydrate, 36.8mg Ammonium Molybdate Tetrahydrate, 5.49g Zinc sulfate monohydrate,
948.2mg ferric chloride hexahydrate)] on, it sets 24 DEG C and cultivates 1 week, [every liter of CMX culture medium includes: 0.1g/ with CM fluid nutrient medium
ML calcium nitrate tetrahydrate solution 10mL, 10mL solution B, 0.5mL trace element solution, 1g yeast extract, 0.5g digest cheese
Element, 0.5g acidolysis casein, 10g glucose, 20g agar powder.(every liter of solution B includes: 20g potassium dihydrogen phosphate, and 25g seven is hydrated
Magnesium sulfate, 15g sodium chloride) (every liter of trace element solution includes: 57.2mg boric acid, 393mg Salzburg vitriol, 13.1mg iodine
Change potassium, 60.4mg Manganous sulfate monohydrate, 36.8mg Ammonium Molybdate Tetrahydrate, 5.49g Zinc sulfate monohydrate, the hydration chlorine of 948.2mg six
Change iron)] scraping, collection spore, micro- sem observation, adjusting spore concentration using haemocytometer is 1 × 106/mL。
B. southern corn leaf blight converts
1mL spore suspension is drawn in 100mLCM fluid nutrient medium, 24 DEG C of shaken cultivation (150rpm) 12-18h, from
The heart collects mycelia and digests 2h in 80mL enzymolysis liquid (3.27g sodium chloride, 0.8g driselase), collects protoplast.By plasm
Body is washed 3 times with 10mLSTC solution, and is finally dissolved in 500 μ LSTC solution (every 100mLSTC solution includes: 21.86g sorb
Alcohol, 1Mtris-HCL1mL, 0.735g CALCIUM CHLORIDE DIHYDRATE) in.The ready PCR fragment of 25mL and 100 μ L protoplasts is molten
Liquid mixes well, addition 1mLPEG solution (it include: polyethylene glycol 30g, 1Mtris-HCL0.5mL in every 50mLPEG solution,
0.37g CALCIUM CHLORIDE DIHYDRATE).It is finally diluted with 1mLSTC solution, and is mixed with regeneration culture medium, 30 DEG C are incubated overnight, each
The bacterium colony that picking extends after water agar 10mL, 30 DEG C of culture 3d of the addition containing 150 μ g/mL hygromycin in culture dish is to containing same
On the CMX culture medium of sample antibiotic.
3) verifying of deletion mutant
Three pairs of primers are selected to screen by PCR amplification to transformant.Amplification meets following result, is determined as
ChCDC3 deletion mutant body: the primer U (5'-CGACAAGAGCGAGTTGAACA- except the homology arm of upstream on genome
It can 3') expand with the primer NLC37 (5'-GGATGCCTCCGCTCGAAGTA-3') of hygromycin gene pairing to expection
The recombinant fragment of size (2.4kb)；Primer D (5'-GGGTGCCATTTGTTGCTATT- except the homology arm of downstream on genome
It can 3') expand with the primer NLC38 (5'-CGTTGCAAGACCTGCCTGAA-3') of hygromycin gene pairing to expection
The recombinant fragment of size (3.0kb)；And code area primers F (5'-ATTGTTGACAACCGCATTCA-3') and R (5'-
CTCCATCTTCTGGAGCTTGG-3') without amplified band (wild-type strain is amplifiable to 0.6kb segment) (see Fig. 3).As a result,
2 plants of ChCDC3 deletion mutant bodies are screened from transformant, are analyzed for follow-up function.
The genetic complement of 3 ChCDC3 deletion mutant body of embodiment
Using primer C-F1 (5'-TCGTGCCTCCTCATTTAACC-3') and C-R1 (5'-CACTGGAACAACTGGCAT
GCACTCCTCGCTCCGAACTAC-3'), expanding southern corn leaf blight ChCDC3 full length gene 2755bp (includes upstream and downstream sequence
Column), using primer C-F2 (5'-CAGGTACACTTGTTTAGAGGTCTCAGATATGGCACCCATGA-3') and C-R2 (5'-
TTGTAGACGACGATGCCGTA-3'), the downstream sequence 827bp of southern corn leaf blight ChCDC3 downstream of gene is expanded.Then with
Carrier p II 99 is template, with DW69 (5'-CATGCCAGTTGTTCCAGTG-3') and DW70 (5'-
It ACCTCTAAACAAGTGTACCTG-3') is II gene of primer amplification npt.Three complementary fragments are transferred to ChCDC3 gene delection
In mutant gene group, using Geneticin as selection markers, genetic complement bacterial strain Δ ChCDC3-C is screened.Select primers F/R into
Row PCR verifying.
Effect of the 4 ChCDC3 gene of embodiment in southern corn leaf blight mycelia growth course
Using plating method, the variation situation of the Relevant phenotypes such as the mycelia growth of ChCDC3 mutant is evaluated.Take 10 μ L
Strain to be tested CMX spore suspension (1 × 106mL-1) it is seeded in the center of solid CMX culture medium, 24 DEG C of cultures, 16h illumination, 8h
It is dark.It has been observed that the colonial morphology of mutant and wild type, complementing strain have significant difference, mutant colonies four after seven days
Zhou Longqi, the speed of growth is obviously slack-off, and thallus lighter, shows that ChCDC3 is southern corn leaf blight mycelia elongation growth
Necessary gene (see Fig. 4,5).
Effect of the 5 ChCDC3 gene of embodiment in terms of southern corn leaf blight conidium generation
Southern corn leaf blight wild-type strain C4, ChCDC3 deletion mutant body and complementing strain are inoculated in respectively solid
On body CMX culture medium, after growth nine days, with 5mL aseptic water washing conidium, spore suspension is collected, blood count is utilized
Plate counts spore, and observes spore shape under the microscope.By compared with wild-type strain, ChCDC3 gene delection
The sporulation quantity of mutant strain is only the 6% of wild-type strain, and conidium yield is decreased obviously (see Fig. 7).By micro-
It has been observed that the conidium shape of ChCDC3 deletion mutant body bacterial strain is irregular, spore diaphragm is reduced, wild-type strain
5-7 diaphragm is usually contained with the conidium of complementing strain, and the conidium of ChCDC3 deletion mutant body bacterial strain is only
There is 1-2 diaphragm (see Fig. 6).By wild-type strain, the sectional growing spore suspension of ChCDC3 deletion mutant body and complementing strain
Liquid is added dropwise respectively on glass slide, and 25 DEG C of moisturizing culture 6h observe conidia germination rate under the microscope.The results show that
The conidia germination rate of ChCDC3 deletion mutant body is only 70% (see Fig. 8) of wild-type strain.These results indicate that
Southern corn leaf blight ChCDC3 gene has important work for southern corn leaf blight illumination, form and germination rate aspect
With.
Effect of the 6 ChCDC3 gene of embodiment in terms of southern corn leaf blight ascospore generation
Southern corn leaf blight wild-type strain C4, ChCDC3 deletion mutant body and complementing strain is small with corn respectively
Pinta bacterium CB7 carries out opposite culture (bacterial strain C4 contains MAT1-1 mating type gene, and CB7 contains MAT1-2 mating type gene).25
DEG C culture 21d after, observe the shell of ascus and ascospore production.It makes discovery from observation, ChCDC3 deletion mutant body
Shell of ascus quantity is only 12% or so of wild-type strain and complementing strain, and shell of ascus yield is decreased obviously (see Fig. 9).Separately
Outside, 7-8 item filiform ascospore is usually contained in the ascus that wild-type strain generates, and ChCDC3 deletion mutant body cannot
Normal ascospore is formed, ascospore is in air bubble-shaped, is easily broken (see Figure 10).It can be seen that southern corn leaf blight
The generation of the ChCDC3 gene pairs shell of ascus and ascospore plays an important role.
Sequence table
Denomination of invention: a kind of southern corn leaf blight ChCDC3 gene and its application
The sequence of SEQ ID No:1
(i) sequence signature: (A) length: 1403 bp；(B) type: nucleotide；(C) chain: single-stranded
(ii) molecule type: DNA
(iii) sequence description: SEQ ID No:1
1 ATGGACCAGA CATTCAACGG GTCCAACCCT GCGGTCCACA CCATCACGGC TAAGAACCCC 61
AAGGCCGCTG CGGCCATGGC CAACGACATG AACATTGTCC GCAGAAAGCT TACCGGCTAT 121
GTTGGTTTCG CCAACCTGCC CAACCAGTGG CACCGCAAGA GTGTGCGCAA GGGATTCAAC 181
TTCAACGTCA TGGTTGTTGG TACGTGTGAC CTGCCAGCAT GCCGTGTCGA ATCTAACTAA 241
CGTGTCGTCC CAGGCGAGTC TGGACTCGGA AAGTCTACCC TTGTAAACAC ACTGTTCAAC 301
ACGTCGCTGT ACCCGCCCAA GGAACGCCAG CCCCCGAGTC TGGACATTTC CAAGACTGTA 361
TCGATCCAGT CCATCAGCGC CGACATTGAG GAGAACGGCG TTCGTCTGAG GTTAACCGTT 421
GTCGATACAC CTGGCTTTGG CGACTTCATC AACAACGATG AGTCCTGGGA CCCCATTGTC 481
AAGAACATTG AGCAACGGTT CGACGCATAC CTGGATGCCG AGAACAAGGT CAACCGCATG 541
AACATTGTTG ACAACCGCAT TCACGCCGTC GTCTACTTTA TTCAGCCTAC GGGCCACTCT 601
CTGAAGCCCA TTGACATTGA GGTGATGAAG AAGCTCCACA CCAAGGTCAA CCTGATTCCC 661
GTCATTGCCA AGGCCGACAC CGTTACGGAC GATGAGATTG ACAACTACAA GAAGAGGGTA 721
AGTAGCTTTA TTTTGCGAAG TGGTTGGGGC TATGTTGACT TGTCCAGATT CTTGCCGACA 781
TTGCGTACCA CAAGATCCAG ATCTTCGAAG GACCCCGGTA TGAGCTTGAC GACGAGGAGA 841
CGATTGCCGA GAACCAGGAG ATCATGGCCA AGGTTCCCTT TGCTGTTGTC GGCTCCAACA 901
CCGAGGTGAC GACGGTTGAC GGCCGCAAGG TCCGCGGACG CGCACTCCCC TGGGGTGTGG 961
TCGAGGTTGA CAACGAGGAG CACTGCGACT TTGTCAAGCT CCGACAGATG CTCATCCGCA 1021
CCCACATGGA GGAGCTCAAG GAAAACACCA ACAATGTCTT GTACGAGAAC TACCGCTCGG 1081
ACAAGCTTGC ACAGATGGGC ATCCAACAGG ACTCGAGCGT GTTCAAGGAG GTCAACCCTG 1141
CTGTCAAGCA AGAGGAGGAG CGCTCGCTGC ACGAGGCCAA GCTCCAGAAG ATGGAGATGG 1201
AGATGAAGAT GGTGTTCCAG CAAAAGGTGC AGGAAAAAGA AAGCAAGCTG CGCCAGAGCG 1261
AGGAAGAGCT GTACGCCCGT CACAAGGAGA TGAAGGACCA GCTGGACCGA CAACGACAGG 1321
AGCTCGAAGA GAAGAAGGCA CGCATCGAGT CTGGCCGGCC GATCGAAGAG AAGGGCAAGC 1381
GAAAGGGCTT CTCGCTCCGC TAA
The sequence of SEQ ID No:2
(i) sequence signature: (A) length: 432 amino acid；(B) type: amino acid；(C) chain: single-stranded
(ii) molecule type: polypeptide
(iii) sequence description: SEQ ID No:2
1 MDQTFNGSNP AVHTITAKNP KAAAAMANDM NIVRRKLTGY VGFANLPNQW HRKSVRKGFN 61
FNVMVVGESG LGKSTLVNTL FNTSLYPPKE RQPPSLDISK TVSIQSISAD IEENGVRLRL 121
TVVDTPGFGD FINNDESWDP IVKNIEQRFD AYLDAENKVN RMNIVDNRIH AVVYFIQPTG 181
HSLKPIDIEV MKKLHTKVNL IPVIAKADTV TDDEIDNYKK RILADIAYHK IQIFEGPRYE 241
LDDEETIAEN QEIMAKVPFA VVGSNTEVTT VDGRKVRGRA LPWGVVEVDN EEHCDFVKLR 301
QMLIRTHMEE LKENTNNVLY ENYRSDKLAQ MGIQQDSSVF KEVNPAVKQE EERSLHEAKL 361
QKMEMEMKMV FQQKVQEKES KLRQSEEELY ARHKEMKDQL DRQRQELEEK KARIESGRPI 421
EEKGKRKGFS LR
Claims (5)
1. a kind of southern corn leaf blight (Cochliobolus heterostrophus) ChCDC3 gene, it is characterised in that: its
DNA sequence dna is as shown in SEQ ID No:1.
2. a kind of protein of southern corn leaf blight (Cochliobolus heterostrophus) ChCDC3 gene coding,
Be characterized in that: its amino acid sequence is as shown in SEQ ID No:2.
3. southern corn leaf blight described in a kind of claim 1 (Cochliobolus heterostrophus) ChCDC3 gene exists
Regulation southern corn leaf blight conidium and ascospore formed in application.
4. a kind of (Cochliobolus heterostrophus) the ChCDC3 gene of southern corn leaf blight described in claim 1 is made
The application in anti-corn southern leaf blight medicament is being designed and screened for target.
5. a kind of (Cochliobolus heterostrophus) the ChCDC3 gene of southern corn leaf blight described in claim 2 is compiled
The protein of code forms the application occurred in defect in southern corn leaf blight conidium and ascospore.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910618994.6A CN110305879B (en) | 2019-07-10 | 2019-07-10 | Maize small leaf spot pathogen ChCDC3 gene and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910618994.6A CN110305879B (en) | 2019-07-10 | 2019-07-10 | Maize small leaf spot pathogen ChCDC3 gene and application thereof |
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| Publication Number | Publication Date |
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